• Journal of Dental Anesthesia and Pain Medicine
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• 제16권1호
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• pp.17-24
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• 2016

Background: To evaluate the antimicrobial activity of lidocaine (LD) topical anesthetic spray against oral microflora. Methods: Antimicrobial effects of 10% LD spray were assessed against six bacterial cultures obtained from volunteers: Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, Streptococcus salivarius, Streptococcus pyogenes, and Streptococcus sanguinis. The filter papers contained $50-{\mu}l$ LD, brain heart infusion (BHI) broth, or 0.2% chlorhexidine. Papers were placed on the cultured blood plates for 1-3 min. After the papers were removed, plates were incubated for 24 h. Bacterial growth on the contact areas was recorded as the antimicrobial score. The split mouth technique was use in for sample collection in clinical study. Filter papers soaked with either BHI broth or LD were placed on the right or left buccal mucosa for 1 min, and replaced with other papers to imprint biofilms onto the contact areas. Papers were placed on blood plates, incubated for 24 h, and antimicrobial scores were determined. Experiments were conducted for 2- and 3-min exposure times with a 1-day washout period. Results: LD exhibited bactericidal effects against E. coli, S. sanguinis, and S. salivarius within 1 min but displayed no effect against S. aureus, E. faecalis, and S. pyogenes. The antimicrobial effect of LD on oral microflora depended upon exposure time, similar to the results obtained from the clinical study (P < 0.05). LD showed 60-95% biofilm reduction on buccal mucosa. Conclusions: Antimicrobial activity of 10% LD topical anesthetic spray was increased by exposure time. The 3 min application reduced oral microflora in the buccal mucosa.

• Asian Pacific Journal of Cancer Prevention
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• 제17권9호
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• pp.4491-4501
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• 2016

Verruco-papillary lesions (VPLs) of the oral cavity described in the literature involve a spectrum of conditions including squamous papilloma, verruca vulgaris, focal epithelial hyperplasia, condyloma, proliferative verrucous leukoplakia and verrucous carcinoma. A majority of the VPLs are slow growing, benign in nature and have a viral aetiology. Virus associated benign mucosal outgrowths are not too difficult to diagnose either clinically or by microscopy. Apart from virus-associated lesions, VPLs harboring malignant potential or behaviour such as verrucous carcinoma, proliferative verrucous leukoplakia, oral verrucous hyperplasia (OVH), oral papillary squamous cell carcinoma (PSCC) and oral conventional squamous cell carcinoma with papillary features (CSCC) need to be further clarified for better understanding of their predictable biologic behavior and appropriate treatment. Current understanding of potentially malignant VPLs is perplexing and is primarily attributed to the use of confusing and unsatisfactory terminology. In particular, the condition referred to as oral verrucous hyperplasia (OVH) poses a major diagnostic challenge. OVH represents a histopathological entity whose clinical features are not well recognised and is usually clinically indistinguishable from a verrucous carcinoma and a PSCC or a CSCC. A consensus report published by an expert working group from South Asia as an outcome of the 'First Asian Regional Meeting on the Terminology and Criteria for Verruco-papillary Lesions of the Oral Cavity' held in Kuala Lumpur, Malaysia, recognised the clinical description of these OVH as a new entity named 'Exophytic Verrucous Hyperplasia'. Previously described clinical features of OVH such as the 'blunt' or 'sharp' variants; and the 'mass' or 'plaque' variants can now collectively fall under this newly described entity. This paper discusses in detail the application of the standardized criteria guidelines of 'Exophytic Verrucous Hyperplasia' as published by the expert group which will enable clinicians and pathologists to uniformly interpret their pool of OVH cases and facilitate a better understanding of OVH malignant potential.

• Advances in Traditional Medicine
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• 제4권3호
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• pp.179-185
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• 2004

The ethanol extract of Indigofera cordifolia was studied for in vivo gastroprotective activity, cytotoxic activity against oral tumor and normal cells, multidrug resistance (MDR) reversal activity, anti-human immunodeficiency virus (HIV) activity and radical scavenging activity. The extract of I. cordifolia showed potent gastric mucosal protective activity against stomach injury induced by HCl/EtOH solution. However, the gastroprotective activity could not be related to the radical mechanism, because the extract weakly scavenged both OH radical and $O_2*^-$. The extract also showed promising levels of MDR-reversing activity. This study demonstrates the tumor-specific cytotoxic action of the plant extract. However, the extract had no anti-HIV activity. From above results, the study suggests the medicinal importance of I. cordiforia extract.

• 대한미생물학회지
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• 제21권1호
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• pp.63-71
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• 1986

The antifungal activities of amphotericin B, 5-fluorocytosine, and ketoconazole in combination of amphotericin B/ketoconazole and 5-fluorocytosine/ketoconazole were determined against 42 strains of Candida spp. isolated from oral cavity. Among 42 strains of Candida species, 36 strains of Candida albicans, 2 strains of Candida parapsilosis and Candida tropicalis 1 strain of Candida krusei and Candida stellatoidea were identified. The minimum inhibitory concentrations(MICs) of amphotericin B, 5-fluorocytosine and ketoconazole for these strains were ranged from 0.05-1.56 mcg/ml, 12.5->100.0 mcg/ml and 0.2-50.0 mcg/ml. In all of the experimental strains, amphotericin B had the greatest antifungal activity on a dilution basis. When a microtiter checkerboard technique was used 5-fluorocytosine acted synergistically with ketoconazole against all strains, whereas amphotericin B has a reduced effect. The killing curve experiments with on strain of Candida albicans WMC-85024 demonstrated that the combination of amphotericin B/ketoconazole and 5-fluorocytosine/ketoconazole produced a decrease in number of colony forming unit of >3 logs in 72 hours.

• 대한미생물학회지
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• 제22권4호
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• pp.403-411
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• 1987

This study investigated whether a correlation exists between proteinase activity, phospholipase activity and adherence of Candida albicans to buccal epithelial cells by using of various strains isolated from oral cavity. The proteinase activity of 30 strains was tested by culture on agar media that contained bovine serum albumin as a nitrogen source. Using the serum-protein-agar method to test proterolysis of serum albumin in 20 strains of Candida albicans. Twenty-six strains of Candida albicans were phospholipase producers and the degree of phopholipase activity of experimental strains were $0.51{\sim}0.89$ measured by Pz-value. Twenty-eight strains of Candida albicans were adhersive to buccal epithelial cells and 15 strains were foung significantly active adherence. Fifteen strains of Candida albicans were correlated with proteinase activity and adherence to epithelial cells and concomitantly 20 strains of Candida albicans were also correlated with phospholipase activity and adherence. In conclusion our investigation provides evidence of a correlation between quantitative proteinase, phospholipase and adherence. An association of these parameters may be an important contributory factor for pathogenicity.

• Journal of Periodontal and Implant Science
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• 제44권2호
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• pp.79-84
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• 2014

Purpose: While single-species biofilms have been studied extensively, we know notably little regarding multispecies biofilms and their interactions. The purpose of this study was to develop and evaluate an in vitro multispecies dental biofilm model that aimed to mimic the environment of chronic periodontitis. Methods: Streptococcus gordonii KN1, Fusobacterium nucleatum ATCC23726, Aggregatibacter actinomycetemcomitans ATCC33384, and Porphyromonas gingivalis ATCC33277 were used for this experiment. The biofilms were grown on 12-well plates with a round glass slip (12 mm in diameter) with a supply of fresh medium. Four different single-species biofilms and multispecies biofilms with the four bacterial strains listed above were prepared. The biofilms were examined with a confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The minimum inhibitory concentrations (MIC) for four different planktonic single-species and multispecies bacteria were determined. The MICs of doxycycline and chlorhexidine for four different single-species biofilms and a multispecies biofilm were also determined. Results: The CLSM and SEM examination revealed that the growth pattern of the multispecies biofilm was similar to those of single-species biofilms. However, the multispecies biofilm became thicker than the single-species biofilms, and networks between bacteria were formed. The MICs of doxycycline and chlorhexidine were higher in the biofilm state than in the planktonic bacteria. The MIC of doxycycline for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis, F. nucleatum, or A. actinomycetemcomitans. The MIC of chlorhexidine for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis or F. nucleatum. Conclusions: To mimic the natural dental biofilm, a multispecies biofilm composed of four bacterial species was grown. The 24-hour multispecies biofilm may be useful as a laboratory dental biofilm model system.

• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3335-3340
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• 2016

Background: Oral submucous fibrosis (OSF) is a precancerous condition with a 4 to13% malignant transformation rate. Related to the habit of areca nut chewing it is mainly prevalent in South-east Asian countries where the habit of betel quid chewing is frequently practised. On chewing, alkaloids and polyphenols are released which undergo nitrosation and give rise to N-nitrosamines which are cytotoxic agents. CYP450 is a microsomal enzyme group which metabolizes various endogenous and exogenous chemicals including those released by areca nut chewing. CYP1A1 plays a central role in metabolic activation of these xenobiotics, whereas CYP2E1 metabolizes nitrosamines and tannins. Polymorphisms in genes that code for these enzymes may alter their expression or function and may therefore affect an individuals susceptibility regarding OSF and oral cancer. The present study was therefore undertaken to investigate the association of polymorphisms in CYP1A1 m2 and CYP2E1 (RsaI/PstI) sites with risk of OSF among areca nut chewers in the Northern India population. A total of 95 histopathologically confirmed cases of OSF with history of areca nut chewing not less than 1 year and 80, age and sex matched controls without any clinical signs and symptoms of OSF with areca nut chewing habit not less than 1 year were enrolled. DNA was extracted from peripheral blood samples and polymorphisms were analyzed by PCR-RFLP method. Gene polymorphism of CYP1A1 at NcoI site was observed to be significantly higher (p = 0.016) in cases of OSF when compared to controls. Association of CYP1A1 gene polymorphism at NcoI site and the risk of OSF (Odd's Ratio = 2.275) was also observed to be significant. However, no such association was observed for the CYP2E1 gene polymorphism (Odd's Ratio = 0.815). Our results suggest that the CYP1A1 gene polymorphism at the NcoI site confers an increased risk for OSF.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2003년도 2003 Annual Meeting, BioExhibition and International Symposium
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• pp.55-62
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• 2003

The mucosal immune system provides a first line of defense against invasion of infectious agents via inhalation, ingestion and sexual contact. For the induction of protective immunity at these invasion sites, one must consider the use of the CMIS, which interconnects inductive tissues, including PP and NALT, and effector tissues of the intestinal, respiratory and genitourinary tracts. In order for the CMIS to induce maximal protective mucosal immunity, co-administration of mucosal adjuvant or use of mucosal antigen delivery vehicle has been shown to be essential. When vaccine antigen is administered via oral or nasal route, antigen-specific Th 1 and Th2 cells, cytotoxic T lymphocytes(CTLs) and IgA B cell responses are effectively induced by the CMIS. In the early stages of induction of mucosal immune response, the uptake of orally or nasally administered antigens is achieved through a unique set of antigen-sampling cells, M cells located in follicle-associated epithelium(FAE) of inductive sites. After successful uptake, the antigens are immediately processed and presented by the underlying DCs for the generation of antigen-specific T cells and IgA committed B cells. These antigen-specific lymphocytes are then home to the distant mucosal effector tissues for the induction of antigen-specific humoral(e.g., IgA) and cell-mediated (e.g., CTL and Th1) immune responses in order to form the first line of defense. Elucidation of the molecular/cellular characteristics of the immunological sequence of mucosal immune response beginning from the antigen sampling and processing/presentation by M cells and mucosal DCs followed by the effector phase with antigen-specific lymphocytes will greatly facilitate the design of a new generation of effective mucosal antigen-specific lymphocytes will greatly facilitate the design of a new generation of a new generation of effective mucosal adjuvants and of a vaccine deliver vehicle that maximizes the use of the CMIS.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1142-1149
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• 2004

Streptococcus mutans has the capacity of inducing dental caries. Thus, to develop a novel way of preventing dental caries, a cell wall hydrolase-producing strain was isolated and its characteristics were investigated. Among 200 alkalophilic strains isolated from soil, 8 strains exhibited lytic activities against Streptococcus mutans. However, strain YU5215 with the highest cell wall hydrolase activity was selected for further study. Strain YU5215 was identified as a novel strain of Bacillus based on analyzing its 16S rDNA sequence and Bergey's Manual of Systematic Bacteriology, and thus designated as Bacillus mutanolyticus YU5215. The optimal conditions for the production of the cell wall hydrolase from Bacillus mutanolyticus YU5215 consisted of glucose ($0.8\%$), yeast extract ($1.2\%$), polypeptone ($0.5\%$), $K_{2}HPO_{4}\;(0.1\%$), $MgSO_{4}{\cdot}7H_{2}O$ ($0.02\%$), and $Na_{2}CO_{3}\;(1.0\%$) at pH 10.0. Bacillus mutanolyticus YU5215 was cultured at 30^{circ}C for 72 h to produce the cell wall hydrolase, which was then purified by acetone precipitation and CM-agarose column chromatography. The molecular weight of the lytic enzyme was determined as 22,700 Da by SDS-PAGE. When the cell wall peptidoglycan of Streptococcus mutans was digested with the lytic enzyme, no increase in the reducing sugars was observed, while the free amino acids increased, indicating that the lytic enzyme had an endopeptidase-like property. The amino terminus of the cell wall peptidoglycan digested by the lytic enzyme was determined as a glutamic acid, while the lytic site of the lytic enzyme in the Streptococcus mutans peptidoglycan was identified as the peptide linkage of L-Ala and D-Glu.

• Journal of Periodontal and Implant Science
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• 제24권3호
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• pp.513-528
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• 1994

배양한 치은섬유아세포와 3T3 세포에서, glycyrrhetinic acid가 cyclosporin A를 처리 세포의 활성에 미치는 영향을 알아보기 위해서 MTT 방법을 이용하여 측정하였다. Cyclosporin A는 $1{\sim}10ng/ml$의 농도에서 인체 치은 섬유아세포와 마우스 3T3 세포 활성을 증가시켰으며, glycyrrhetinic acid는 농도와 비례하게 마우스 3T3 세포의 성장을 억제시켰다. 특히 $25{\mu}g/ml$ 이상의 농도에서는 현저하게 억제시킴을 관찰할 수 있었다. 반면, 인체 치은 섬유아세포에서는 $50{\mu}g/ml$ 이상의 농도에서 조차도 성장을 억제 시키지 않았다. 또한, 마우스 3T3와 인체 치은 섬유아세포에서 일정한 cyclosporin A 농도에 $1-100{\mu}g/ml$의 다양한 농도로 glycyrrhetinic acid를 첨가하였을 때, cyclosporin A 단독으로 처리한 세포에 비하여 유의성 있게 세포의 활성을 억제시켰으며, 특히, $100{\mu}g/ml$의 glycyrrhetinic acid를 첨가하였을 경우 세포활성을 현저하게 억제시켰다. 3T3 세포에 cyclosporin A와 $50{\mu}g/ml$의 glycyrrhetinic acid를 함께 처리한 경우 glycyrrhetinic acid단독처리한 군에 비하여 세포활성에 첨가 효과를 보였으며, 인체 치은 섬유아세포 에서는 같이 처리한 경우 glycyrrhetinic acid단독 처리에 비하여 뚜렷한 억제 효과를 나타내었다. 이러한 상승효과는 1ng/ml 의 cyclosporin A와 $50{\mu}g/ml$ glycyrrhetinic acid를 같이 처리한 군에서 가장 뚜렷하게 관찰되었다. 이러한 결과로, 인체 치은 섬유아세포에 cyclosporin A와 glycyrrhetinic acid를 동에 처리하였을 경우 세포의 활성에 상승 효과가 있음을 알 수 있다.